Rabbit anti gata3

ILC2 cells were lysed by cold RIPA buffer (87787, Thermo Fisher Scientific, Waltham, MA, USA). Total protein lysates (25 μg/lane) were separated via 12% SDS-PAGE and then transferred to polyvinylidene fluoride membranes. The membranes were subsequently blocked with 5% nonfat milk in Tris-Buffered Saline and Tween-20 (TBST, maintaining 20 mM Tris-HCl, 0.15 M NaCl, 0.05% Tween-20, pH 7.5) for 2 h at room temperature and further incubated overnight with rabbit anti-Hes1 (1:500, Abcam, Cambridge, MA, USA), rabbit anti-GATA3 (1:500, Abcam, Cambridge, MA, USA), rabbit anti-NF-κB (1:500), rabbit anti- Notch (1:500, Abcam, Cambridge, MA, USA), rabbit anti- RORα (1:1000, Abcam, Cambridge, MA, USA), and mouse anti-β-actin antibodies (1:800, Abcam, Cambridge, MA, USA) at 4°C according to the manufacturer’s instructions. After washing with Tris-Buffered Saline and Tween 20 (TBST, Beyotime Biotech. Shanghai, China) for three times, the membranes were incubated with the secondary antibody (Abcam, Cambridge, MA, USA) and finally processed using an enhanced chemiluminescence (ECL) reaction kit (Cell Signaling Technology, Danvers, MA, USA). The relative band densities of the target proteins relative to the β-actin band density were quantified using the Bio-Rad Quantity One 1-D analysis software package (Bio-Rad, Hercules, CA, USA).